Functional verification of Related Genes Susceptible to DENV2 in Aedes aegypti

The dengue virus (DENV) is predominantly transmitted by Aedes aegypti. Investigating genes associated with mosquito susceptibility to DENV2 offers a theoretical foundation for targetedinterventions to regulate or block viral replication and transmission within mosquitoes. Based on the transcriptomic analyses of the midgut and salivary glands from Aedes aegypti infected with DENV2, alongside analyses of Aag2 cell infections, 24 genes potentially related to the regulation of Aedes aegypti infection with DENV2 were identified. To assess their roles, transient transfection, overexpression models in Aag2 cells, and mosquito gene knockdown models were established.Changes in viral load before and after gene manipulation were compared to evaluate the effects of differentially expressed genes (DEGs) on viral replication. Overexpression of 19 DEGs in Aag2 cells resulted in significant changes in DENV2 RNA copies in the cell supernatant (P < 0.05). In adultmosquitoes, knockdown of defensin-A, defensin-A-like, and SMCT1 reduced DENV2 RNA copies, while knockdown of UGT2B1 and ND4 increased them. These findings highlight the roles of defensin-A, defensin-A-like, SMCT1, UGT2B1, and ND4 in modulating DENV2 replication, offering valuable insights into mosquito-virus interactions.